Microscope optical system, microscope, and virtual slide forming system

ABSTRACT

An observation field of view of a microscope can be moved without moving or changing an objective lens and without varying position or state of a sample. A microscope optical system has a mirror that changes the direction of the optical path by reflection and is located in the optical path between an objective lens of the microscope and an image to be observed. The mirror is able to be tilted by changing the position of a reflecting surface of the mirror. Accordingly, the observation field of view is moved by tilting the mirror. In other words, the observation field of view can be moved without changing a positional relationship between the objective lens of the microscope and the sample.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a division of application Ser. No. 11/121,961 filedMay 5, 2005 now U.S. Pat. No. 7,630,113. The disclosure of the followingpriority application is herein incorporated by reference:

Japanese Patent Application No. 2004-140264 filed on May 10, 2004.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to a microscope and an optical systemthereof and a virtual slide forming system and in particular to atechnique for moving an observing field of view in a microscope.

2. Related Background Art

In ordinary microscope observation, after searching for a point to beobserved with an objective lens having low magnification, the point isobserved with enlarging by an objective lens having high magnification.However, there is a possibility of danger that a position of a sample isslipped out by an impact upon changing an objective lens.

In an example disclosed in Japanese Patent Application Laid-Open No.2002-31758, in order to observe a wide area, the magnification of theobjective lens is limited to 32 or less and to observe details with highmagnification, the numerical aperture of the objective lens is set to0.85 or more. And the objective lens and a magnification-change opticalsystem is combined. Accordingly, without changing the objective lens, itbecomes possible to observe a wide field of view by setting lowmagnification to the magnification-change optical system and a finedetail with high resolving power by setting high magnification to themagnification-change optical system.

However, in the method disclosed in Japanese Patent ApplicationLaid-Open No. 2002-31758, the same position of a sample always comes tothe center of the observing image whenever changing the objective lens.Accordingly, in a low magnification observation, when you find aposition, which you feel like to observe with enlarging, locating awayfrom the center of the field of view, in order to observe with highmagnification, you have to bring the position near to the optical axisof the objective lens. Accordingly, the positional relation between thesample and the objective lens has to be changed, in other words, theobserving field of view has to be moved.

In an ordinary upright microscope, when an observing field of view is tobe moved, it has been carried out that the observer moves a stage onwhich the sample is placed perpendicularly to the optical axis of theobjective lens or moves the objective lens.

However, in a microscope observation in a field such aselectrophysiology, it may be happened that a manipulator or the like isset to a sample, so that great care has to be taken that the manipulatordoes not touch with the objective lens upon moving the observation fieldof view. Moreover, it also has to be careful that the objective lensdoes not touch with a tube or a channel arranged for maintaining theenvironment of the sample.

In an actual experiment system, the distance between the tip of theobjective lens and the manipulator is extremely small (1 mm or less), sothat it has been very difficult to observe without touching with eachother. Moreover, when observing with an immersion-objective lens, it mayhappen that the interface between the objective lens and the water,being in the state of contact with each other, makes a change by movingthe field of view. In this case, when the sample is vulnerable to thevibration, there is a possibility of danger that the state of contactbetween an electrode needle set on the tip of the manipulator and thesample or the sample itself is changed.

Accordingly, it has been demanded to provide a method to move theobservation field of view without moving or changing the objective lensor without changing the position of the stage or the sample.

SUMMARY OF THE INVENTION

The present invention is made in view of the aforementioned problems andhas an object to provide a technique capable of moving an observationfield of view of a microscope without changing the position or the stateof a sample and a stage and without moving or changing of an objectivelens.

According to a first aspect of the present invention, a microscopeoptical system includes a first objective lens and a second objectivelens. The first objective lens locates at the most sample side in anoptical path between a sample and an image to be observed. The secondobjective lens forms an intermediate image conjugate with the sampletogether with the first objective lens. The microscope optical systemaccording to the first aspect of the present invention has specialfeatures described below. Firstly, a mirror that changes the directionof the optical path by reflection and locates in the optical pathbetween the first objective lens and the second objective lens isincluded. Secondly, the mirror can be tilted with changing the positionof a reflecting surface of the mirror.

In the microscope optical system according to the first aspect of thepresent invention, it is preferable that a variable magnificationoptical system capable of changing magnification of the image to beobserved is included between the mirror and the image to be observed.

In the microscope optical system according to the first aspect of thepresent invention, the microscope optical system preferably includes acontroller that has information showing relations between a position ofan observation field of view corresponding to the sample and an amountof tilt of the mirror and tilts the mirror on the basis of theinformation.

A microscope according to a second aspect of the present invention hasthe microscope optical system according to the first aspect of thepresent invention.

A third aspect of the present invention provides a virtual slide formingsystem including the microscope according to the second aspect, an imagedata generator and an image synthesizer. The image data generatorrepeatedly carries out in the microscope a movement that tilts the tiltmirror and a movement that photographs the image to be observed andgenerates image data. The image synthesizer synthesizes a plurality ofthe generated image data and forms virtual slides of the sample.

According to a fourth aspect of the present invention, a microscopeoptical system including a first objective lens, and a second objectivelens is provided. The first objective lens locates at the most sampleside in an optical path between a sample and an image to be observed.The second objective lens forms an intermediate image conjugate with thesample together with the first objective lens. The microscope opticalsystem according to the fourth aspect of the present invention includesa lens that locates in the optical path behind the intermediate imageand is movable in the direction perpendicular to the optical axis.

A fifth aspect of the present invention provides a microscope equippedwith the microscope optical system according to the fourth aspect of thepresent invention.

A sixth aspect of the present invention provides a virtual slide formingsystem that includes the microscope according to the fifth aspect of thepresent invention, an image data generator, and an image synthesizer.The image data generator repeatedly carries out in the microscope amovement that moves the lens in a direction perpendicular to the opticalaxis and a movement that photographs the image to be observed andgenerates image data. The image synthesizer synthesizes a plurality ofthe generated image data and forms virtual slides of the sample.

A seventh aspect of the present invention provides a microscope opticalsystem including a first objective lens, a mirror, and a secondobjective lens. The first objective lens locates at a sample side in anoptical path between a sample and an image to be observed. The mirrorlocates to an image side of the first objective lens. The secondobjective lens locates to the image side of the mirror and forms anintermediate image conjugate with the sample. The mirror is arranged tobe able to be tilted such that the intermediate image is moved in thedirection substantially perpendicularly to the optical axis.

In the microscope optical system according to the seventh aspect of thepresent invention, the microscope optical system preferably includes acontroller that has information showing relations between a position ofan observation field of view corresponding to the sample and an amountof tilt of the mirror and tilts the mirror on the basis of theinformation.

An eighth aspect of the present invention provides a microscopeincluding the microscope optical system according to the seventh aspectof the present invention.

A ninth aspect of the present invention provides a virtual slide formingsystem including the microscope according to the eighth aspect of thepresent invention, an image data generator and an image synthesizer. Theimage data generator repeatedly carries out in the microscope a movementthat tilts the tilt mirror and a movement that photographs the image tobe observed and generates image data. The image synthesizer synthesizesa plurality of the generated image data and forms virtual slides of thesample.

Other features and advantages according to the present invention will bereadily under stood from the detailed description of the preferredembodiments in conjunction with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic diagram showing a microscope system according to afirst embodiment of the present invention.

FIG. 2 is a schematic diagram showing the microscope of FIG. 1 indetail.

FIG. 3 is a schematic plan view of a variable magnification opticalsystem in detail seen along the optical axis of the objective lens shownin FIG. 2 in the direction from an eyepiece portion to a sample.

FIG. 4 is a diagram explaining the optical path of a bundle of lightcome out from the objective lens forming an image in an observationcamera through the variable magnification optical system according tothe first embodiment.

FIG. 5 is a diagram explaining a relation of observation fields of viewbetween with low magnification and with high magnification.

FIG. 6 is a schematic diagram showing a microscope system according to asecond embodiment of the present invention.

FIG. 7 is a schematic diagram showing a microscope system according to athird embodiment of the present invention.

FIG. 8 is a schematic diagram showing the microscope of FIG. 7 indetail.

FIG. 9 is a schematic plan view of a fixed optical system 116 and amoving optical system 120 shown in FIG. 8 in detail seen along theoptical axis of the objective lens shown in FIG. 8 in the direction froman eyepiece portion to a sample.

FIG. 10 is a diagram explaining the optical path of a bundle of lightcome out from the objective lens forming an image in a camera throughthe variable magnification optical system according to the thirdembodiment.

FIG. 11 is a schematic diagram showing a microscope system (a virtualslide forming system) according to a fourth embodiment of the presentinvention.

FIG. 12 is a perspective view showing a large stage of the microscopeshown in FIG. 11 in detail.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

Each embodiment according to the present invention is explained belowwith reference to accompanying drawings. In each drawing, the samereference symbol is attached to the same element so as to omitduplicated explanation.

First Embodiment

FIG. 1 is a schematic diagram showing a microscope system according to afirst embodiment of the present invention. As shown in FIG. 1, themicroscope system 10 has a microscope 12, a mirror inclinationcontroller 16, a computer 18, and a monitor 20. The computer 18 obtainsan image signal from a camera 24 of the microscope 12 through a cable 26and displays an enlarged image of a sample on the monitor 20.

FIG. 2 is a schematic diagram showing the microscope 12 of FIG. 1 indetail. As shown in FIG. 2, the microscope 12 has a tilt mirror 28, aneyepiece portion 32, the camera 24, a variable magnification opticalsystem 36, a fluorescence excitation light source 38, an objective lens40, a stage 44, and a transmitted illumination light source 46.

The tilt mirror 28 is a semi-transparent mirror or a dichroic mirror.Accordingly, a bundle of light transmitting through the tilt mirror 28is formed an observing image by an optical system (not shown because ofpublicly known elements) such as an eyepiece in the eyepiece portion 32.Incidentally, when the system does not have the eyepiece portion 32, thetilt mirror 28 may be a total reflection mirror. The fluorescenceexcitation light source 38 illuminates a sample 50 on the stage 44downward (epi-illumination) through such as lenses and mirrors (notshown). The transmitted illumination light source 46 illuminates asample 50 on the stage 44 upward through such as lenses and mirrors (notshown).

The objective lens 40 according to the first embodiment of the presentinvention has a magnification of 16 and a numerical aperture of 0.8 andmakes it possible to observe a wide field of view. The objective lens 40has, in order from the sample 50 side, a first lens group, a second lensgroup, and a third lens group (not shown). The first lens group includesa meniscus lens having a concave surface facing to the sample 50 sideand has positive refractive power as a whole. The second lens groupincludes at least two three-piece cemented lenses. The third lens groupincludes a first cemented meniscus lens having a concave surface facingto an image side, and a second cemented meniscus lens having a concavesurface facing to the sample 50 side and facing to the concave surfaceof the first cemented meniscus lens.

The objective lens 40 according to the first embodiment of the presentinvention satisfies the following conditional expressions (1) through(4):0.23<d0/F<0.35  (1)1<f1/F<2  (2)0.7<r3g/F<1.0  (3)1.6<n3g<1.75  (4)where d0 denotes a distance along the optical axis between the sampleplane and the most sample 50 side surface of the objective lens 40, Fdenotes the focal length of the objective lens 40, f1 denotes the focallength of the first lens group, r3g denotes a radius of curvature of theconcave surface of the first cemented meniscus lens in the third lensgroup, and n3g denotes refractive index at d-line (wavelength λ=587.6nm) of the image side lens composing the first cemented lens in thethird lens group.

When the ratio d0/F is equal to or exceeds the upper limit ofconditional expression (1), the bundle of light extends largely, so thatit becomes difficult to make the numerical aperture large. On the otherhand, when the ratio d0/F is equal to or falls below the lower limit ofconditional expression (1), it becomes difficult to secure sufficientworking distance. When the ratio f1/F is equal to or exceeds the upperlimit of conditional expression (2), refractive power of the first lensgroup becomes weak, so that it becomes difficult to bring the bundle oflight sufficiently near to parallel. On the other hand, when the ratiof1/F is equal to or falls below the lower limit of conditionalexpression (2), spherical aberration and coma become worse. When theratio r3g/F is equal to or exceeds the upper limit of conditionalexpression (3), sufficient effect for correcting Petzval sum cannot beobtained, so that flatness of the image plane is deteriorated bycurvature of field and astigmatism. On the other hand, when the ratior3g/F is equal to or falls below the lower limit of conditionalexpression (3), the radius of curvature becomes too small, so thatcorrection of coma and the like is affected. When the value n3g is equalto or exceeds the upper limit of conditional expression (4),transmittance of the light in short wavelength range becomes low. On theother hand, when the value n3g is equal to or falls below the lowerlimit of conditional expression (4), it becomes difficult to correctspherical aberration and coma.

FIG. 3 is a schematic plan view of a variable magnification opticalsystem 36 of the microscope 12 in detail seen along the optical axis ofthe objective lens 40 shown in FIG. 2 in the direction from an eyepieceportion 32 to the sample 50. As shown in FIG. 3, the variablemagnification optical system 36 includes a low magnification opticalsystem 54, a high magnification optical system 56, and a mirror 58. Thelow magnification optical system 54 includes variable magnificationlenses 60 and 62 and has a magnification of 0.35. The high magnificationoptical system 56 includes variable magnification. lenses 64 and 66 andhas a magnification of 4.

In other words, when either the low magnification optical system 54 orthe high magnification optical system 56 is inserted on the optical pathbetween the tilt mirror 28 and the camera 24, the magnification of theobserving image projected on the camera 24 can be changed. Accordingly,the magnification of the observing image is determined by the product ofthe magnification (16) of the objective lens 40 multiplied by that ofthe variable magnification optical system 36, and, in this embodiment,is 5.6 and 64.

FIG. 4 is a diagram explaining the optical path of a bundle of lightcome out from the objective lens 40 forming an image in the camera 24through the variable magnification optical system 36 according to thefirst embodiment. The principle of moving method of the observationfield of view in the microscope system 10 is explained below withreference to FIG. 4.

A bundle of light from the sample 50, after passing the objective lens40, is reflected by the tilt mirror 28 changing the direction of theoptical path, and is incident to the variable magnification opticalsystem 36. After passing the variable magnification lens 64 (or 60), thebundle of light forms an intermediate image 37, after passing throughthe variable magnification lens 66 (or 62), changes the optical pathperpendicularly by the mirror 58, and is incident to an imaging lens 68in the camera 24. By the imaging lens 68, an enlarged image of thesample 50 is formed on the imaging plane of an imaging device 69 in thecamera 24. The camera 24 carries out publicly known processing and thelike to the image signal output from the imaging device 69 and generatesimage data. The image data is input to the computer 18 through the cable26. By the way, the image processing and the like may be carried out bythe computer 18 other than by the camera 24. By the way, a field stopmay be arranged in the vicinity of the intermediate image 37.

In the first embodiment, the mirror inclination controller 16 shown inFIG. 1 controls a tilt controller 70 on the tilt mirror 28 to tilt thetilt mirror 28 in response to the instruction from the computer 18. Thetilt in this case means such that the existing position of thereflection surface of the tilt mirror 28 changes by the tilt, and doesnot mean that the reflection surface is rotated in the same plane aroundthe normal to the reflection surface.

By the way, the position and the inclination of the mirror 58 shown inFIG. 4 are fixed. The angle between the reflection surface of the mirror58 and the optical axis of the variable magnification optical system 36is 45 degrees. In the first embodiment, even if the variablemagnification optical system 36 is changed to either the lowmagnification optical system or the high magnification optical systemshown by an arrow in FIG. 3, the position of the optical axis is notchanged. In the microscope according to the first embodiment, opticalarrangement changes only when the tilt mirror 28 is tilted.

The tilt mirror 28 has two axes of tilt both of which are located on thereflection surface of the tilt mirror 28. By the way, the both axes areimaginary axes, it means that the tilt mirror 28 tilts around theimaginary axis, and not that a shaft like a stick or bar is installed.In order to define the direction of the axis of tilt of the tilt mirror28, a normal state where the reflection surface of the tilt mirror 28 isparallel to that of the mirror 58 is assumed. In the normal state, thebundle of light come out from the objective lens 40 changes the opticalpath perpendicularly by the tilt mirror 28.

In the normal state, one of the axes of the tilt mirror 28 isperpendicular to both the axis of the objective lens 40 and that of thevariable magnification lens 64 and 66 (or 60 and 62), in other words, inthe direction perpendicular to the plane of FIG. 4. Hereinafter, theaxis is called as an X axis. The other tilt axis of the tilt mirror 28crosses the X axis at right angles on the reflection surface of the tiltmirror 28 and is hereinafter called as a Y axis. The cross point of theX axis and the Y axis is located at the center of the reflection surfaceof the tilt mirror 28 and the tilt mirror 28 is tilted without changingthe position of the cross point.

When the tilt mirror 28 is tilted, the intermediate image 37 formed bythe objective lens 40 and the variable magnification lens 64 (or 60)moves in the same plane perpendicular to the optical axis of thevariable magnification optical system 36. Accordingly, the bundle oflight incident to the variable magnification lens 66 (or 62) locatedbehind the intermediate image 37 on the optical path moves in thedirection perpendicular to the optical axis of the variablemagnification optical system 36. This is equivalent to the case that thestage 44 on which the sample 50 is placed is moved perpendicularly tothe optical axis of the objective lens 40. In other words, a user canmove the observation field of view displayed on the monitor 20 bytilting the tilt mirror 28 with input operation to the computer 18.Incidentally, the direction of the bundle of the light passing throughthe tilt mirror 28 does not change in spite of tilting the tilt mirror28, so that the same image is observed by the eyepiece portion 32.

FIG. 5 is a diagram explaining a relation of observation fields of viewbetween with low magnification 501 and with high magnification 502. Thespecific method for moving the observation field of view is explainedbelow with reference to FIG. 5. In the first embodiment of the presentinvention, for the purpose of simplify the explanation, it is explainedwith assuming that the movement of the observation field by tilting thetilt mirror 28 is not carried out upon observing with low magnification.In other words, it is explained the case that after searching for aposition to be observed with low magnification observation, by changingthe variable magnification optical system 36 to high magnification, theenlarged position is observed with controlling the tilt mirror 28.

As shown in FIG. 5, in the observation with low magnification 501, theblock separation (dotted lines) of the field of view and the coordinatesof the field of view are superimposed on an observing image on themonitor 20 and each block is assumed to be a unit block field of view503. By the way, it is possible that only the observing image is simplydisplayed. In FIG. 5, the coordinates and the dotted lines of the blockfield of view are shown only a portion to avoid complication.

In order to specifically explain the amount of tilt (tilt angle) of thetilt mirror 28, which becomes important information in moving theobservation field of view, optical value of each element is shown as anexample. The aspect ratio of the picture on the monitor 20 of thehorizontal direction (right and left in FIG. 5) to the verticaldirection (up and down in FIG. 5), in other words, the aspect ratio ofthe imaging device 69 is 4:3. The diagonal dimension of the imagingdevice 69 is a two-third inch.

Accordingly, when the variable magnification optical system 36 is lowmagnification, the total magnification of the microscope 12 is 5.6, sothat the diameter of about 1.6 mm on the sample 50 becomes theobservation field of view. When the variable magnification opticalsystem 36 is high magnification, the total magnification of themicroscope 12 is 64, so that the diameter of about 130 μm on the sample50 becomes the observation field of view.

In order to simplify the explanation, the imaging device 69 and the likeare assumed to be arranged such that when the tilt mirror 28 is tiltedaround the X axis, the field of view moves only up or down, and when thetilt mirror 28 is tilted around the Y axis, the field of view moves onlyright or left. Moreover, when the variable magnification optical system36 is changed to high magnification without tilting the tilt mirror 28,the block field of view of the coordinates (0, 0) locating at the centerof the observing image in the low magnification is assumed to be thestandard block. The observing image of the standard block with highmagnification is shown by 502 in FIG. 5 on the right side.

In order to observe enlarged image of a block adjacent upward ordownward to the standard block, in other words, the block (0, 1) or (0,−1), the tilt mirror 28 may be tilted by about 0.2970° around the X axisand the variable magnification optical system 36 may be changed to thehigh magnification. In order to move to the block field of view of twoblocks upward, the tilt amount may be 0.594° and in order to move to theblock field of view of n-blocks upward (or downward), the tilt amountmay be n×0.297°.

In order to observe enlarged image of a block adjacent right or left tothe standard block, in other words, the block (−1, 0) or (1, 0), thetilt mirror 28 may be tilted by about 0.3210° around the Y axis and thevariable magnification optical system 36 may be changed to the highmagnification. In order to move to the block field of view of n-blocksright (or left), the tilt amount may be n×0.321°. The above-describedtilt amount can be controlled by constructing the tilt controller 70 bya micro-actuator, a linear actuator with an encoder, or the like. By theway, the operation for moving the field of view may be carried out afterchanging the variable magnification optical system 36 to highmagnification.

The computer 18 stores the table data showing that how much amountshould the tilt mirror 28 be tilted around the X axis and the Y axis inorder to observe each block field of view in the observation picture oflow magnification with high magnification. A portion of the table datais, for example, shown below.

Coordinates (0, 0):[θx (0, 0), θy (0, 0)]

Coordinates (0, 1):[θx (0, 1), θy (0, 1)]

Coordinates (1, 1):[θx (1, 1), θy (1, 1)]

As shown in the first embodiment, when the tilt mirror 28 is set to thestandard state upon observing with low magnification, θx (0, 0) and θy(0, 0) are both 0°. In this case, θx (0, 1) is 0°, θy (0, 1) is 0.321°,θx (1, 0) is 0.297°, and θy (1, 0) is 0°. Accordingly, when a blockfield of view is selected by inputting with a keyboard, the computer 18derives the tilt amount of the tilt mirror 28 to move to the selectedblock field of view in accordance with the table data. The computer 18instructs the mirror inclination controller 16 to tilt the tilt mirror28 by the derived tilt amount. The mirror inclination controller 16controls the tilt controller 70 to tilt the tilt mirror 28. Then, thevariable magnification optical system 36 is changed to highmagnification by a publicly known mechanism in response to theinstruction by the computer 18.

When the tilt amount of the tilt mirror 28 becomes large, it may happenthat the relation between the moving amount of the observing field ofview and the tilt amount of the tilt mirror 28 is not linear. In thiscase, when the field of view is simply moved by a multiple of the eachmoving amount of a unit block field of view, a distorted quadranglefield of view on the sample 50 is observed as an area of a rectangularshape in the image plane. Accordingly, each moving amount in the tabledata according to the first embodiment is a value corrected thedistortion, so that there is some difference from the valuecorresponding to a multiple of the moving amount of each block field ofview. By correcting distortion as described above, always a rectangulararea on the sample 50 can be observed. The tilt amount corrected thedistortion can be calculated from experiments, simulations, or the likein advance.

In this manner, the first embodiment makes it possible to move theobservation field of view by only changing the amount of tilt of thetilt mirror 28 inserted into the collimated optical path between theobjective lens 40 and the imaging lens 68. In other words, moving theobservation field of view does not require a change or a move of theobjective lens 40 or a move of the stage 44. Accordingly, an inevitableproblem of the prior art that the positional relation between theobjective lens and the sample has to be changed upon moving theobservation field of view has been solved.

Moreover, even if a needle of a micromanipulator is there in thevicinity of the objective lens 40, there is no danger of contact. Sincethe operation is just selecting the block field of view to the computer18, observation field can be moved with an extremely simple operation.As a result, convenience of a user is greatly improved.

Since only the tilt mirror 28 is moved upon moving the observation fieldof view, there is no possibility to have an influence such as vibrationon the sample 50, so that there is no possibility to vary the state ofthe sample 50. Since just tilting the tilt mirror 28, the observationfield of view can be moved with a high speed. Furthermore, byquantitatively controlling the tilt mirror 28, the positional state ofthe tilt mirror 28 upon selecting the same block field of view can bethe same regardless of the number of moving times of the observationfield of view.

Accordingly, when the same block field of view is selected, the sameobservation picture of the previous observation can be displayed withhigh reproducibility without producing a positional gap of the tiltmirror 28. In this case, since the table data defining the tilt angle ofthe tilt mirror 28 is corrected with respect to the above-mentioneddistortion, the observation can always be carried out by moving on arectangular area on the sample 50.

When observation magnification is changed without tilting the tiltmirror 28, the center of the observation field of view with lowmagnification corresponds with that with high magnification. By usingthis function, a plurality of positions to be observed are registeredwith their coordinates as the center of the observation field of view inlow magnification in advance, and the coordinates of each point aredesignated in turn after changing to high magnification, so thateffective observation can be carried out.

In the first embodiment of the present invention, it is explained anexample in which observation field of view is moved stepwise with eachblock field of view. However, the present invention is not limited tothis embodiment. By operating such as a lever connected to the computer18, the observation field of view may be moved continuously in thedirection that the lever is inclined. Alternatively, the observationfield of view may be moved stepwise with superposing half block field ofview. This is the same as the third embodiment explained later.

It is explained an example that when the tilt mirror is tilted aroundthe X axis, the field of view is moved only upward or downward, and whenthe tilt mirror is tilted around the Y axis, the field of view is movedonly right or left, and vice versa. Alternatively, when the mirror istilted around either X axis or Y axis, the field of view may be set tomove both upward or downward and right or left. In this case also, whenthe table data such as the one described above has been prepared inadvance, the observation field of view can be moved to any position.

The method to derive the tilt angle of the tilt mirror 28 by thecomputer 18 need not to use the above-described table data. For example,the tilt angle of the tilt mirror 28 may be derived from a conversionexpression.

In order to simplify the explanation, although an example is explainedthat the position of the intersection of the axes of the tilt mirror 28is fixed, another example may be possible that the position of theintersection of the axes of the tilt mirror 28 is moved.

In order to simplify the explanation, although the observation field ofview is not moved by the tilt mirror 28 upon observing with lowmagnification, it may be moved.

In the first embodiment, it is explained an example that the mirror (thetilt mirror 28) locating to the objective lens 40 side is tilted and themirror (mirror 58) locating to the observer side is fixed. The presentinvention is not limited to this embodiment. The observation field ofview may be moved by tilting the mirror locating to the observer sidewith fixing the mirror locating to the objective lens side.

Second Embodiment

FIG. 6 is a schematic diagram showing a microscope system according to asecond embodiment of the present invention. As shown in FIG. 6, amicroscope system 80 according to the second embodiment includes amanipulator controller 84 and a micromanipulator 86 in addition to theelements of the microscope system 10 according to the first embodiment.The difference between the first embodiment and the second embodiment isthat movement of the observation field of view is associated withmovement of the micromanipulator 86 such that when the observation fieldof view is moved to the right, an electrode needle of themicromanipulator 86 is also moved to the right.

The difference between the second embodiment and the first embodiment isspecifically explained below. For example, the instruction that theobservation field of view is moved in association with themicromanipulator 86 is assumed to be input to the computer 18. In thiscase, when the instruction to move the observation field of view isinput to the computer 18, the computer 18 instructs the inclinationcontroller 16 to tilt the tilt mirror 28 as same as the firstembodiment.

Synchronizing with this movement, on the basis of the tilt angle of thetilt mirror 28, the computer 18 derives whether in which direction andby which amount the observation field of view moves on the sample 50, inother words, the moving amount and the moving direction of theobservation field of view on the sample 50. Then, the computer 18instructs the manipulator controller 84 to change the position of themicromanipulator 86 in accordance with the derived moving direction andmoving amount.

As described above, the second embodiment makes it possible toautomatically synchronize the position of the micromanipulator 86 to theobservation field of view upon moving the observation field of view.Accordingly, the operational burden to a user can be lightened relativeto the case that movement of the observation field of view and theoperation of the micromanipulator 86 are carried out separately.Moreover, the micromanipulator 86 can be operated with moving theobservation field of view. Furthermore, a plurality of micromanipulators86 are set to the sample 50, the movement of the observation field ofview can be synchronized with the positions of the plurality ofmicromanipulators 86. In the second embodiment also, since amechanically moving element is only the tilt mirror 28, the similareffect as the first embodiment can be obtained.

Third Embodiment

FIG. 7 is a schematic diagram showing a microscope system according to athird embodiment of the present invention. As shown in FIG. 7, amicroscope system 100 includes the monitor 20, the computer 18, thecable 26, a cable 108, the manipulator controller 84, themicromanipulator 86, and a microscope 110 according to the thirdembodiment.

FIG. 8 is a schematic diagram showing the microscope 110 of FIG. 7 indetail. As shown in FIG. 8, the microscope 110 includes the eyepieceportion 32, a fixed mirror 114, a fixed optical system 116, a movingoptical system 120 (including the camera 24), the fluorescenceexcitation light source 38, the objective lens 40, the stage 44, and thetransmitted illumination light source 46.

The difference between the third embodiment and the second embodiment isa mechanism for moving the observation field of view in the microscope110. In the third embodiment, the observation field of view is moved bymoving the moving optical system 120. Moreover, in the third embodiment,the inclination or the position of the fixed mirror 114 locating on theoptical axis of the objective lens 40 is not changed. The fixed mirror114 is a semi-transparent mirror or a dichroic mirror.

FIG. 9 is a schematic plan view of the fixed optical system 116 and themoving optical system 120 shown in FIG. 8 in detail seen along theoptical axis of the objective lens shown in FIG. 8 in the direction fromthe eyepiece portion 32 to the sample 50. As shown in FIG. 9, the fixedoptical system 116 includes variable magnification lenses 130 and 132.The moving optical system 120 includes a relay optical system 136 andthe mirror 58. The relay optical system includes a variablemagnification lenses 138 and 140. As shown in FIG. 10 explained later,although the moving optical system 120 includes a moving mechanism 148,the imaging lens 68, and the camera 24, they are omitted in FIG. 9 toavoid complication.

In the third embodiment, the variable magnification optical system 36 iscomposed of the fixed optical system 116 and the relay optical system136. In other words, the variable magnification optical system 36 is setto low magnification (for example, 0.35) by inserting the variablemagnification lens 130 of the fixed optical system 116 and the variablemagnification lens 138 of the relay optical system 136 into the opticalpath between the fixed mirror 114 and the mirror 58. The variablemagnification optical system 36 is set to high magnification (forexample, 4) by inserting the variable magnification lens 132 of thefixed optical system 116 and the variable magnification lens 140 of therelay optical system 136 into the optical path between the fixed mirror114 and the mirror 58. The magnification of the microscope 110 is givenby the product of the magnification of the variable magnificationoptical system 36 multiplied by that of the objective lens 40 and lowmagnification and high magnification can be changed automatically asshown in the first embodiment.

FIG. 10 is a diagram explaining the optical path of a bundle of lightcome out from the objective lens 40 forming an image in the camera 24according to the third embodiment. The principle of moving theobservation field of view in the microscope 100 is explained below withreference to FIG. 10.

The bundle of light from the sample 50 through the objective lens 40 isreflected by the fixed mirror 114 changing the optical pathperpendicularly, and forms an intermediate image 37 after passingthrough the variable magnification lens 130 (or 132). Then, the bundleof light passes through the variable magnification lens 138 (or 140), isreflected by the mirror 58 changing the optical path perpendicularly,passes through the imaging lens 68, and forms an observing image on thedetection surface of the imaging device 69. By the way, a field stop maybe arranged in the vicinity of the intermediate image 37.

In the third embodiment, upon moving the observation field of view, themoving optical system 120 as a whole is moved perpendicularly to theoptical axis of the variable magnification optical system 36. In otherwords, the relay optical system 136, the imaging lens 68, and the camera24 are moved in a body, and each element of the moving optical system120 is moved on the same plane. Since the movement is on the same plane,there are two moving directions, one is, for example, in the directionof the optical axis of the imaging lens 68. In this case, the other isin the direction perpendicular to both the optical axis of the imaginglens 68 and that of the variable magnification lens, in other words, inthe direction perpendicular to the plane of FIG. 10.

For example, when the moving optical system 120 is moved downward alongthe optical axis of the imaging lens 68, the position of each element isshown by dotted line in FIG. 10. In this case, when a bundle of lightforming the central part of the intermediate image 37 is incident to thevariable magnification lens 138 (or 140) before moving, a bundle oflight forming a part of the intermediate image 37 away from the centeris incident to the variable magnification lens 138 (or 140) aftermoving. This is equivalent to the case that the stage 44 on which thesample 50 is placed is moved perpendicularly to the optical axis of theobjective lens 40. Accordingly, in the third embodiment also, theobservation field of view can be moved without changing positionalrelation between the objective lens 40 and the sample 50.

Then, the procedure of moving the observation field of view isexplained. In order to simplify the explanation as same as the firstembodiment, it is explained the case that after searching for a positionto be observed with low magnification observation, by changing thevariable magnification optical system to high magnification, theenlarged position is observed with moving the moving optical system 120.

As same as the first embodiment, the instruction to move the observationfield of view is carried out, for example, by selecting the block fieldof view with inputting from the keyboard of the computer 18. Thecomputer 18 stores “the table data that defines the each moving amountof the moving optical system 120 in the above-described two directionsin order to move the observation field of view to each block field ofview”.

When a block field of view is selected, the computer 18 derives themoving amount of the moving optical system 120 in the above-describedtwo directions on the basis of the table data. The moving mechanism 148is connected to the computer 18 through the cable 108. The computer 18controls the moving mechanism 148 to move the moving optical system 120in accordance with the derived moving amount, and moves the observationfield of view. Then, the variable magnification optical system 36 may bechanged to high magnification.

In the third embodiment also, the same effect as the first and secondembodiments can be obtained. By the way, in the third embodiment, anexample is explained that the microscope system 100 includes themanipulator controller 84 and the micromanipulator 86. Althoughduplicated explanation is omitted, an example is explained that movementof the observation field of view and movement of the micromanipulator 86are synchronized. However, the present invention is not limited to theembodiment. As same as the first embodiment, the construction may notinclude the manipulator controller 84 or the micromanipulator 86.

In order to simplify the explanation, although the observation field ofview is not moved by the moving optical system 120 upon observing withlow magnification, it may be moved. The method to derive the movingamount of the moving optical system 120 by the computer 18 need not touse the above-described table data, and it may be derived from aconversion expression.

Fourth Embodiment

FIG. 11 is a schematic diagram showing a microscope system 160 accordingto a fourth embodiment of the present invention. As shown in FIG. 11,the microscope system 160 includes the monitor 20, the computer 18, thecable 26, a cable 170, the mirror inclination controller 16, and amicroscope 174 according to the fourth embodiment.

The microscope 174 includes the fluorescence excitation light source 38,the camera 24, the variable magnification optical system 36, theeyepiece portion 32, the tilt mirror 28, the tilt controller 70, theobjective lens 40, a large stage 180, a stage moving mechanism 184, andthe transmitted illumination light source 46. The microscope 174 is thesame as the microscope 12 according to the first embodiment excepthaving the large stage 180 and the stage moving mechanism 184 instead ofthe stage 44. The stage moving mechanism 184 is connected to thecomputer 18 through the cable 170.

FIG. 12 is a perspective view showing a large stage 180 of themicroscope 174 shown in FIG. 11 in detail. As shown in FIG. 12, aplurality of sample slides 190 are placed on the large stage 180.

In the fourth embodiment, the computer 18 tilts the tilt mirror 28stepwise without moving the large stage 180 in a state where thevariable magnification optical system 36 is set to high magnification.Accordingly, the computer 18 moves stepwise the sample area shown as theimage (in other words, a position on the sample slide 190 whose image isformed on the imaging device 69 is moved stepwise). This is similar tothe movement of the observation field of view according to the firstembodiment. However, the purpose is not observation but to form virtualslides.

Then, the computer 18 obtains image data generated by the camera 24. Thecomputer 18 displays a picture image from the obtained image data on themonitor 20 and stores the data in the memory.

The computer 18 repeatedly carries out the movements that tilts the tiltmirror 28 (moving the sample area), instructs the camera 24 to generateimage data, and obtains and stores the image data. In this manner, thecomputer 18 connects the image data generated successively to form thewhole image data of a single sample slide 190 (virtual slide). Thecomputer 18 stores the generated virtual slide in such as a hard disk.

Then, the computer 18 controls the stage moving mechanism 184 to movethe large stage 180 in the direction perpendicular to the optical axisof the objective lens 40 (for example, the direction indicated by anarrow shown in FIG. 12), and to locate another sample slide 190 directlyunder the optical axis of the objective lens 40. As described above, thecomputer 18 forms a virtual slide of the sample slide 190 and stores itin such as a hard disk. The above-described explanation is the operationof the fourth embodiment.

It frequently happens that a virtual slide is formed collectively to alarge number of large sample slides (such as a 25 mm square), so that itis necessary to use a large stage in order to place these sample slides.Moreover, since high resolving power is required, it is necessary tophotograph with high magnification, a high numerical aperture (such as0.65 or more), and a large number of pixels (such as 2000 or morevertically, and 2500 or more horizontally). Accordingly, in order toform virtual slides quickly, it is necessary to move the stage or theobjective lens a large number of times with high speed. In this case, itis highly possible that the sample is affected by the vibration and thelike. Accordingly, the stage has not been possible so far to be movedwith high speed upon forming virtual slides, so that it has been desiredto make it possible.

Accordingly, in the fourth embodiment, the method according to the firstembodiment is applied to forming virtual slides so that the movement ofthe sample area shown by the image is precisely controlled with highspeed by the computer 18, the tilt mirror 28, and the like. As a result,the picture image composing each part of the virtual slides canprecisely be photographed with high speed. Therefore, theabove-described conventional problems can be solved, and the time toform the virtual slides can be shortened.

In the virtual slide, since the image data of the whole sample slidesare formed by connecting each image data photographed with moving thefield of view, it is necessary that direction of each image data isprecisely aligned with each other. In order to satisfy the requirement,it is desirable that the tilt amount of the tilt mirror should becorrected to correct a minute amount of image rotation caused by themirror (in this case, the tilt mirror 28). The correction is derived bysuch as experiments or simulation in advance, and may be shown in thetable data.

In the fourth embodiment, it is explained an example that the movementof the sample area shown by the image is moved by tilting the tiltmirror 28 with adding the large stage 180 and the stage moving mechanism184 to the microscope 12 according to the first embodiment. However, thepresent invention is not limited to the fourth embodiment. With addingthe large stage 180 and the stage moving mechanism 184 to the microscope110 according to the third embodiment, the similar operation may becarried out by moving the moving optical system 120. The movement andphotographing of the sample area shown by the image may be carried outin the state that the variable magnification optical system 36 is set tolow magnification. In this case, the resolving power of the formedvirtual slide is lowered.

In the first through fourth embodiment, it is explained an example thatthe magnification of the variable magnification optical system 36 ischanged by changing lenses. The present invention is not limited to theembodiment. By constructing the variable magnification optical system 36by lens groups capable of zooming, the magnification of the variablemagnification optical system 36 may be changed without changing thelens.

In the first through the third embodiment, although it is explained anexample that an upright microscope is applied to the present invention,an inverted microscope may also be applied to the present invention.

All numerical values listed in the first through fourth embodiment areonly an example for reference except those connected with the objectivelens 40.

As described above, the present invention can considerably be used inthe field of microscope (in particular, a microscope forelectrophysiology or virtual slides).

Additional advantages and modification will readily occur to thoseskilled in the art. Therefore, the invention in its broader aspect isnot limited to the specific details, and representative devices shownand described herein. Accordingly, various modifications may be madewithout departing from the spirit or scope of the general inventiveconcept as defined by the appended claims and their equivalents.

1. A microscope optical system comprising: an objective lens to belocated nearest to a sample in an optical path between the sample and animage to be observed; a variable magnification optical system that formsan intermediate image conjugate with the sample, together with theobjective lens; a mirror having a reflecting surface disposed in theoptical path between the objective lens and the variable magnificationoptical system, the mirror being arranged to change position of thereflecting surface by tilting of the mirror so as to change a directionof the optical path; a mirror controller that controls the tilting ofthe mirror; a micromanipulator that manipulates the sample; amanipulator controller that controls movement of the micromanipulator;and a computer operatively connected to the mirror controller and themanipulator controller, wherein the computer instructs the mirrorcontroller to tilt the mirror and simultaneously therewith determines adirection and an amount of the displacement of the observation field ofview based on an angle of the tilt of the mirror, and instructs themanipulator controller to change a position of said micromanipulatorbased on the determined direction and the amount of the displacement ofthe observation field of view.
 2. A microscope optical system accordingto claim 1, further comprising a plurality of said micromanipulators. 3.A microscope optical system according to claim 1, further comprising: acamera arranged so as to receive light flux which has passed through theobjective lens, been reflected by the mirror, and been transmittedthrough the variable magnification optical system, wherein the camera isconnected to the computer, and the direction and the amount of thedisplacement of the observation field of view is determined based on animage obtained by the camera.
 4. The microscope optical system accordingto claim 1, wherein the mirror controller controls tilting of the mirrorso as to change the observation field of view without moving theobjective lens and the sample.
 5. A microscope optical systemcomprising: an objective lens to be located nearest to a sample in anoptical path between the sample and an image to be observed; a variablemagnification optical system that forms an intermediate image conjugatewith the sample, together with the objective lens; a mirror having areflective surface disposed in the optical path between the objectivelens and the variable magnification optical system, the mirror beingarranged to change position of the reflecting surface by the tilting ofthe mirror so as to change a direction of the optical path; a mirrorcontroller that controls the tilting of the mirror; a micromanipulatorthat manipulates the sample; a manipulator controller that controlsmovement of the micromanipulator; and a computer operatively connectedto the mirror controller and the manipulator controller, wherein thecomputer instructs the mirror controller to tilt the mirror andinstructs the manipulator controller to change a position of saidmicromanipulator based on an angle of the tilt of the mirror.
 6. Themicroscope optical system according to claim 5, wherein the mirrorcontroller controls tilting of the mirror so as to change theobservation field of view without moving the objective lens and thesample.